pBM062 vector (V016191)

LOCUS       62056_4135        9926 bp DNA     circular SYN 28-JUN-2019
DEFINITION  Cloning vector pBM062, complete sequence.
ACCESSION   MN019114
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9926)
  AUTHORS   Markus BM, Bell GW, Lorenzi HA, Lourido S.
  TITLE     Optimizing Systems for Cas9 Expression in Toxoplasma gondii
  JOURNAL   mSphere 4 (3), e00386-19 (2019)
  PUBMED    31243081
REFERENCE   2  (bases 1 to 9926)
  AUTHORS   Markus BM.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JUN-2019) Whitehead Institute, 455 Main Street, 
            Cambridge, MA 02142, United States
REFERENCE   3  (bases 1 to 9926)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "mSphere"; 
            date: "2019"; volume: "4"; issue: "3"; pages: "e00386-19"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-JUN-2019) Whitehead Institute, 455 Main Street, Cambridge, MA 
            02142, United States"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     5'UTR           651..1193
                     /label=TUB1-cat-SaG1
                     /note="TUB1-cat-SaG1"
     CDS             1200..1853
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     3'UTR           1857..2185
                     /label=SAG1
                     /note="SAG1"
     misc_feature    2197..2696
                     /label=U6 upstream region
                     /note="U6 upstream region"
     misc_feature    2697..2717
                     /label=sgRNA-1 protospacer
                     /note="sgRNA-1 protospacer"
     misc_RNA        2718..2793
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2794..2808
                     /label=PolIII terminator
                     /note="PolIII terminator"
     5'UTR           2821..3319
                     /label=TgTUB1
                     /note="TgTUB1"
     misc_feature    3324..3389
                     /note="3xFLAG; three tandem FLAG(R) epitope tags, followed
                     by an enterokinase cleavage site"
     misc_feature    3396..3416
                     /note="SV40 NLS; nuclear localization signal of SV40 large
                     T antigen"
     CDS             3441..7541
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     misc_feature    7542..7589
                     /note="nucleoplasmin NLS; bipartite nuclear localization
                     signal from nucleoplasmin"
     polyA_signal    7623..7830
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7839..7979
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      8054..8509
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8791..8895
                     /label=AmpR promoter
     CDS             8896..9753
                     /label=AmpR
                     /note="beta-lactamase"